Method for manufacturing red mold dioscorea

ABSTRACT

The present invention discloses a method for manufacturing red mold dioscorea, and the method comprises the following steps: washing and cutting a fresh dioscorea into pieces with a specific dimension; drying the pieces of the fresh dioscorea for making the dried dioscorea contain a specific water content and a specific sulfur content; adding some water with an appropriate ratio to fresh dioscorea or dried dioscorea; sterilizing the dioscorea; after the dioscorea being cooled down, inoculating the dioscorea with  Monascus  species; cultivating the dioscorea with an appropriate temperature, an appropriate humidity and an appropriate shacking frequency for an appropriate time period; and drying the cultivated red mold dioscorea with an appropriate water content.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to a method for manufacturing red molddioscorea, and more particularly relates to a method for manufacturingred mold dioscorea with a solid state cultivation method or a liquidstate cultivation method.

2. Description of the Prior Art

Hypertension caused by hypercholesterol is a popular civilizationdisease in recent years. Following the advancement of medical science,populations of middle-aged and geriatric people are increasing. It isdifficult to eliminate superoxide free radicals in middle-aged andgeriatric people result from the lower metabolic mechanism, and themiddle-aged and geriatric diseases include cancer and cardiopathy areprone to occur. Cardiovascular diseases arise from hypercholesterolemiainclude apoplexy, coronary atherosclerotic cardiopathy and hypertension.In pursue of the healthy diet in life, health food nowadays is verypopular. Red mold products possess both effects of inhibition ofcholesterol synthesis and blood pressure lowering and become one of thepopular health food.

Numerous researches in recent years prove that red mold rice havesignificant lipid-lowering effect on hyperlipidemia hamsters, thus redmold rice have potential to be an ingredient in cholesterol-loweringdrugs and improve this kind of disease. The above effects are resultfrom that red mold rice can produce several kinds of secondarymetabolite, such as cholesterol-synthesis inhibitor, blood pressurelowering material, anti-cancer material and so forth. Some previousresearches about secondary metabolite of red mold yeast indicate thatthe yield and pattern of secondary metabolite may be directly orindirectly influenced by cultural environment and method.

Recently, red mold related products made by using some specificfermented substrate are proposed. In 2006, Lee et al. propose theirstudy of “Monascus fermentation of dioscorea for increasing the productof cholesterol-lowing agent”, and the study is then published on Journalof Applied Microbial and Cell Physiology. In Lee's study,Monascus-fermented substrate, for example, Monascus fermented dioscorea,is proven its effect on increasing the production of monaclin K andmonascin, wherein the monaclin K can be used as a cholesterol-loweringagent and the monascin can be as a anti-inflammation agent.

Please refer to FIG. 1, there is shown a flowchart of a red moldsubstrate preparation method proposed by Lee. As shown in FIG. 1, toprepare the red mold substrate, a substrate is firstly purchased fromlocal supermarket (S01′). Then, five grams of substrate is soaked indistilled water for 8 hr (S02′), and excess water is removed by using asieve (S03′). After the steps of S01′˜S03′ are finished, the substrateis then autoclaved in a wood dish at 120° C. for 20 min (S04′). Andafter being cooled, the substrate is inoculated with a 5% sporesuspension (10⁷ spores/ml) and 0.3% ethanol (S05′). Therefore, theinoculated substrate is cultivated at 30° C. for 10 days (S06′);moreover, during the culturing stage, 145 ml of water is added onceevery 12 hr at a total of three times and the addition of water startson the fifth day of cultivation. Therefore, Lee successfully stimulatesthe formation of monacolin K by adopting above-mentioned preparationmethod. In addition, Lee also finds that the monascus-fermenteddioscorea have more amount of yellow pigment than monascus-fermentedrice. Yellow pigment of Monascus species including monascin andankaflavin has been shown as an anticancer agent and anti-inflammationagent.

Therefore, because Lee's preparation method is proven the effect onincreasing the production of monaclin K and monascin, the preparationmethod is then widely used for the production of monaclin K andmonascin, and called solid state cultivation method. However, in spiteof Lee's preparation method is helpful to the production of monaclin Kand monascin, his preparation method still includes the shortcomings anddrawbacks as follows:

1. In the steps S02′ and S03′ of Lee's preparation method, the substrateis soaked in distilled water for 8 hr and the excess water is removed byusing a sieve. The person skilled in this art is able to find that Leestill fails to optimize a best water ratio for soaking the substrate,and the water ration of steps S02′ and S03′ is obviously unsuitable tothe sterilization of the substrate.2. In Lee's preparation method, it does not provide a suitableenvironment for culturing the inoculated substrate, for example, asuitable environment with an optimal relative humidity. Watersupplementation is used in Lee's preparation method, but the environmenthumanity control is more useful to stabilize the production of monacolinK, monascin, ankaflavin, and citrinin, the person skilled in this artknows that.3. Although Lee's preparation method is proven the effect on increasingthe production of monaclin K and monascin can be increased, it alsohelpful to increase the production of citrinin, which is grouped into akind of polyketide derivatives of Monascus. So that Lee adds 0.3% (v/w)ethanol to substrate (dioscorea) in order to lower citrinin production.However, the procedure also significantly decreased the production ofthe formational metabolites monascin and ankaflavin by 2.5 and 3-fold.

In addition, differing from the solid state cultivation method forincreasing the production of monaclin K and monascin, in 2007, Leefurther proposes his study of “Improving the ratio of Monacolin K tocitrinin production of Monascus purpureus NTU 568 under dioscorea mediumthrough the mediation of pH value and ethanol addition”, and the studyis then published on Journal of Agricultural and Food Chemistry. In thisstudy, Lee proposes a liquid state cultivation method for increasing theproduction of monaclin K and monascin. Please refer to FIG. 2, whichillustrates a flowchart of the liquid state cultivation method proposedby Lee. As shown in FIG. 2, to prepare and culture themonascus-fermented dioscorea (substrate), a dry dioscorea root isfirstly milled into powder (60-80 mesh) and used as the substrate ofseed culture (S01″), and a culture medium with 5% the powdered dioscoreais then prepared (S02″). After the steps of S02′˜S02′ are finished, theculture medium is subsequently inoculated with 5% Monascus sporesolution and 0.3% ethanol (S03″). Therefore, in step S04″, thefermentation is carried out with aeration rate at 4 vvm, 30° C. and theagitation at 200 rpm for 12 days (S04″). After fermentation, thedioscorea is collected and dried at 55° C. for 48 hr (505″), andeventually to weight the total dry biomass for the calculation of theratio of dry biomass weight to dioscorea added weight (B/D ratio)(S06″).

Through the liquid state cultivation method, Lee successfully stimulatesthe formation of monacolin K and monascin. Besides, Lee also determinesan optimum culture conditions based on experiment results. However, inspite of Lee's liquid state cultivation method is helpful to theproduction of monaclin K and monascin, his method still includes themain drawback of “lacking citrinin-inhibiting step”.

So that, in view of the above-mentioned conventional red mold substratepreparation method (including the solid state cultivation method and theliquid state cultivation method) still have shortcomings and drawbacks,it is necessary to choose a fermented substrate with better effects andadjust the culture conditions to be optimum so as to increase theproduction of monaclin K, monascin and ankaflavin, and then to elevatethe effects of blood-lipid lowering, blood-pressure lowering andatherosclerosis prevention. Therefore, the inventor of the presentinvention resorted to past experience, imagination, and creativity,performed experiments and researches repeatedly, and eventually devisedthe present invention: a method for manufacturing red mold dioscorea.

SUMMARY OF THE INVENTION

The first objective of the present invention is to provide a method formanufacturing red mold dioscorea, owing to the amounts of each kind ofsecondary metabolite produced by red mold yeast are variant underdifferent cultural conditions, the manufacturing method and all thecultural conditions according to the present invention can promote theamount of formation of effective secondary metabolites, such asanti-inflammation material (monascin), anti-cancer material (ankaflavin)and cholesterol-lowering material (monacolin K), which can be used forcompositions of drugs for lowering blood lipid, lowering blood pressureand preventing atherosclerosis.

Accordingly, in order to achieve the above-mentioned objectives of thepresent invention, the inventors propose a method for manufacturing redmold dioscorea, comprising the steps of:

(1) washing a fresh dioscorea clean and cutting the fresh dioscorea intopieces with a specific dimension, wherein the specific dimension is 2˜20mm;(2) drying the pieces of the fresh dioscorea for making the drieddioscorea contain a specific water content and a specific sulfur contentby a sulfuring method, wherein the specific water content is below 15%;(3) adding a distilled water to the dried dioscorea for making the drieddioscorea and the water be a specific ratio in volume, wherein thespecific ratio is 1:0.75 and this specific ratio make the dioscoreasuitable to be executed a sterilization process;(4) soaking the dried dioscorea in the distilled water for a specifictime period;(5) proceeding the sterilization process of the dioscorea at 121° C. for20 min;(6) spreading the steamed dioscorea in a wood dish for cooling thesteamed dioscorea;(7) inoculating the dioscorea with 5% Monascus spore solution;(8) cultivating the inoculated dioscorea at a specific temperature and aspecific cultural humidity for a specific cultural time period, whereinthe specific cultural temperature is 30° C., the specific culturalhumidity is 60%, and the specific cultural time period is 10 days;(9) proceeding an anaerobic treatment to the product of the step (8) fora specific treating time period, wherein the anaerobic treatment iscarried out by culturing the inoculated dioscorea under 100% CO₂condition; and(10) drying the product of the step (9) to a specific water content.

Moreover, for achieving the above-mentioned objectives of the presentinvention, the inventors propose a second embodiment of the method formanufacturing red mold dioscorea, comprising the steps of:

(1) adding a specific amount of water to a raw material of a dioscoreato make the raw material of the dioscorea and the water be a specificratio, wherein raw material of the dioscorea is a dried dioscoreacontaining sulfur, and the sulfur contained by the dried dioscorea maycontribute an acidic condition with pH 3-4, and the acidic conditionbeing able to simulate the production of secondary metabolites of thered mold dioscorea when the red mold dioscorea is cultured;(2) proceeding a sterilization process to the raw material of thedioscorea and then cooling the product down to a specific temperature;(3) inoculating a Monascus spp. to the sterilized dioscorea;(4) culturing the inoculated dioscorea under a specific culturaltemperature and a specific shaking frequency for a specific culturaltime period;(5) proceeding an anaerobic treatment to the product of step (4) for aspecific treating time period, wherein the anaerobic treatment iscarried out by culturing the inoculated dioscorea under 100% CO₂condition; and(6) drying the product of step (5) to a specific water content, whereinthe specific water content is below 15%.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a flowchart of a red mold substrate preparation methodproposed by Lee;

FIG. 2 is a flowchart of a liquid state cultivation method proposed byLee;

FIG. 3A and FIG. 3B are a flowchart of a method for manufacturing redmold dioscorea according to a first preferred embodiment of the presentinvention;

FIG. 4 is a flowchart of the method for manufacturing the red molddioscorea according to a second preferred embodiment of the presentinvention;

FIG. 5 is a trend graph of yellow pigment and red pigment formation inthe cultural processes of the red mold rice and red mold dioscorea; and

FIG. 6 is a trend graph of monacolin K formation in the culturalprocesses of the red mold rice and red mold dioscorea.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

To achieve the foregoing objectives and effects, the inventors integratered mold yeast with dioscorea and improve and amend the conditions ofmanufacturing methods, thus achieving the method for manufacturing redmold dioscorea of the present invention. Hereinafter, flowcharts ofmethods for manufacturing red mold dioscorea according to a firstpreferred embodiment and a second preferred embodiment of the presentinvention will be described in detail for illustrating the method formanufacturing red mold dioscorea.

Referring to FIG. 3A and FIG. 3B, there are shown a flowchart of amethod for manufacturing red mold dioscorea according to the firstpreferred embodiment of the present invention. As shown in FIGS. 3A and3B, to execute the method for making a red mold dioscorea, it is firstlywashing a fresh dioscorea clean and cutting the fresh dioscorea intopieces with a specific dimension (step S01), in which the specificdimension is 2˜20 mm and the optimal dimension is 2 mm. The method isnext to proceed step S02, drying the pieces of the fresh dioscorea formaking the dried dioscorea contain a specific water content and aspecific sulfur content by a sulfuring method, wherein the specificwater content is below 15% and specific sulfur content is below 160 ppm.After step S02 is completed, step S03 is then proceeded for adding adistilled water to the dried dioscorea, so as to make the drieddioscorea and the water be a specific ratio in volume, wherein thespecific ratio is 1:0.75 and this specific ratio make the dioscoreasuitable to be executed a sterilization process.

Next, the method is proceeded steps S04 and S05 to soak the drieddioscorea in the distilled water for 60 minute and execute thesterilization process of the dioscorea at 121° C. for 20 min. And StepS06 is then executed in order to spread the steamed dioscorea in a wooddish for cooling the steamed dioscorea. So that, after the steameddioscorea is cooled, the method next proceeds to step S07 forinoculating the dioscorea with 5% Monascus spore solution. The cultivatestep, i.e., step S08, which is subsequently executed, such that theinoculated dioscorea is cultivated at a specific temperature and aspecific cultural humidity for a specific cultural time period, whereinthe specific cultural temperature is 30° C., the specific culturalhumidity is 60%, and the specific cultural time period is 10 days.

Continuously, the method is proceeded steps S10 in order to proceed ananaerobic treatment to the product of the step S08 for a specifictreating time period, wherein the anaerobic treatment is carried out byculturing the inoculated dioscorea under 100% CO₂ condition, and thespecific treating time period is 48 hr. Eventually, step S10 is executedfor drying the product of the step S09 to a specific water content,wherein the specific water content is below 15%.

In recent years, the secondary metabolites of red mold yeast are paidmore and more attention by people, wherein monascin and ankaflavin, theyellow pigments, are proved to be the anti-inflammation agents and theactive ingredients for lowering the incidence of cancer, and monacolin Kis an effective cholesterol-lowering material, thus promoting thecontents of monascin, ankaflavin and monacolin K in the Monascusfermented products is recently an important goal for Monascusresearches.

In the past, red mold related products are all made from rice as thefermented substrate, but the yields of anti-inflammation material(monascin), anti-cancer material (ankaflavin) and cholesterol-loweringmaterial (monacolin K) are not abundant in red mold rice. In order toincrease the amounts of these active ingredients, Lee et al. propose asolid state cultivation method for increasing the production of monaclinK and monascin. However, Lee's method includes the shortcomings anddrawbacks of (1) failing to optimize a best water ratio for soaking thesubstrate; (2) does not provide a suitable environment for culturing theinoculated substrate; and (3) adding 0.3% (v/w) ethanol to substrate(dioscorea) in order to lower the production of citrinin, one kind ofpolyketide derivatives of Monascus.

The aforesaid first preferred embodiment of the method introduced by thepresent invention is also a solid state cultivation method forincreasing the production of the secondary metabolites of red mold, suchas monacolin K, monascin and ankaflavin. In the method of the presentinvention, dioscorea is soaked in a distilled water with the drieddioscorea and the water having a specific ratio (1:0.75) in volume, andthis optimal ratio is suitable to the sterilization process of thedioscorea. Moreover, when the inoculated dioscorea is cultivated, thesulfurized dioscorea fabricated by the step S02 is able to stimulate theproduction of monascin and ankaflavin by 1.5 and 1.3 times via an acidiccondition (pH 3-4) contributed from S₂. In addition, in the method ofthe present invention, the 60% of controlled environment humanity isuseful to stabilize the production of monacolin K, monascin andankaflavin, and such environment humanity control is easier to beapplied on a large industrial scale than water supplementation.

Furthermore, in order to effectively inhibit the production of citrininbut keep the production of monacolin K, monascin and ankaflavin, ananaerobic treatment is adopted in the method of the present inventionand carried out by culturing the inoculated dioscorea under 100% CO₂condition. In the solid fermentation of the present invention, anaerobetreatment can obviously decrease the citrinin concentration by 2.1times. Therefore, according to above descriptions of the first preferredembodiment of the method for manufacturing red mold dioscorea, itbelieves that the red mold dioscorea can be manufactured successfully byutilizing the method of the present invention.

Please refer to FIG. 4, which illustrates a flowchart of a method formanufacturing red mold dioscorea according to the second preferredembodiment of the present invention. The method for manufacturing redmold dioscorea of the present invention not only includes the firstpreferred embodiment but also has a second preferred embodiment; thesecond preferred embodiment is a liquid state cultivation method. Asshown in FIG. 4, to execute the second preferred embodiment, it isfirstly adding a specific amount of water to a raw material of adioscorea to make the raw material of the dioscorea and the water be aspecific ratio (step S01 a), wherein the specific ratio is ranged from1:10 to 1:200, and the raw material of the dioscorea is a drieddioscorea with sulfur below 15%, so as to keep the sulfur content isbelow 160 ppm. In the method of second preferred embodiment, the sulfurcontained by the dried dioscorea may contribute an acidic condition withpH 3-4, and the acidic condition is able to simulate the production ofsecondary metabolites of the red mold dioscorea when the red molddioscorea is cultured.

Next, the method proceeds to step S02 a for proceeding a sterilizationprocess to the raw material of the dioscorea at 121° C. for 30 minutes,and then cooling the product down to a specific temperature. After thedioscorea is cooled, step S03 a is subsequently executed for inoculatinga Monascus spp. to the sterilized dioscorea. Then, in step S04 a, itcultures the inoculated dioscorea under a specific cultural temperatureand a specific shaking frequency for a specific cultural time period,wherein the specific cultural temperature in the step S04 a is 25˜37°C., the specific shaking frequency is 50˜300 rpm, and the specificcultural time period is 8˜20 days. During the culture, in order toinhibit the production of citrinin, it needs to proceeding an anaerobictreatment to the cultured dioscorea for 48 hr, and n the anaerobictreatment is carried out by culturing the inoculated dioscorea under100% CO₂ condition. Therefore, after the anaerobic treatment isfinished, the step S06 a is then executed for drying theMonascus-fermented dioscorea with the water content, below 15%.

Therefore, Comparing to the conventional liquid state cultivation methodproposed by Lee et al., the introduced liquid state cultivation methodaccording to the second preferred embodiment of the present inventionincludes the main advantage as following: citrinin-inhibiting step isadopted in the liquid state cultivation method of the present invention,the citrinin-inhibiting step is carried out by culturing the inoculateddioscorea under 100% CO₂ condition. Moreover, it needs to especiallynote that this citrinin-inhibiting step (i.e., anaerobic treatment)would not simultaneously decease the production of the secondarymetabolites of the Monascus-fermented dioscorea.

According to above descriptions, it believes that red mold dioscorea(i.e., Monascus-fermented dioscorea) can be manufactured successfullyaccording to the method of the present invention. There are largedifferences in the appearances between red mold dioscorea and red moldrice, wherein the largest distinction is that the major pigment producedby red mold rice is red pigment and the appearance of red mol dioscoreais orange-yellow.

Referring to FIG. 5, which is a trend graph of yellow pigment and redpigment formation in the cultural processes of red mold rice (solidcircle) and red mold dioscorea (hollow circle). A large amount of yellowpigment is produced by red mold dioscorea in the initial stage, and theyield of yellow pigment in red mold dioscorea is higher than that in redmold rice; in terms of red pigment, the yield of red pigment in red molddioscorea is lower than that in red mold rice. These results demonstratethat red mold yeast fermented with dioscorea can produce a large amountof yellow pigment containing some active ingredients, such as monascinand ankaflavin.

Referring to the following table 1, which compares the yield ofmonascin, ankaflavin and GABA in red mold dioscorea and red mold ricecultured with different methods.

TABLE 1 Monascin (mg/kg) Ankaflavin (mg/kg) GABA (mg/kg) Red mold rice -3547 1598 131 solid state cultivation Red mold dioscorea - 15011 10074513 solid state cultivation Red mold rice - 5415 2488 45 liquid statecultivation Red mold dioscorea - 23280 15330 46 liquid state cultivation

As shown in the table 1, the yield of monascin in red mold dioscorearesulted from solid state cultivation is 4.23 times more than that inred mold rice resulted from solid state cultivation, and the yield ofankaflavin in red mold dioscorea resulted from solid state cultivationis elevated by 6.30 times significantly contrasting with red mold rice.This shows that the contents of monascin and ankaflavin in red molddioscorea cultured by solid state cultivation is higher than that in redmold rice cultured by solid state cultivation. Red mold dioscoreaproduced by liquid state cultivation has the same effects as aboveresults. The contents of monascin and ankaflavin in red mold dioscoreacultured by liquid state cultivation are 4.30 and 5.16 timesrespectively more than that in red mold rice cultured by liquid statecultivation. Furthermore, the production quantity of GABA is also higherin the cultural products in red mold dioscorea.

Referring to FIG. 6, which is a trend graph of monacolin K formation inthe cultural processes of red mold rice (solid circle) and red molddioscorea (hollow circle). The trend graph illustrates that the slowproduction of monacolin K in red mold rice begins at the second day ofthe cultivation and stagnates at the sixth day; red mold dioscoreaproduces monacolin K constantly until the tenth day, wherein theproduction of monacolin K is very rapid from the fifth day to the tenthday. Additionally, after completing the cultivation, the yield ofmonacolin K in red mold dioscorea is over 5 times more than that in redmold rice.

Referring to the following table 2, which compares that the yields ofmonacolin K, red pigment and yellow pigment in several kinds ofsubstrates.

TABLE 2 Monacolin K Red pigment (mg/kg) (A500/g) Yellow pigment (A400/g)dioscorea 2584 ± 127 30 ± 5.0 74 ± 4.7 rice 481 ± 33 50 ± 4.2 41 ± 4.8cassava 522 ± 34 50 ± 3.8 34 ± 3.8 Sweet potato 196 ± 21 51 ± 3.2 39 ±3.9 potato 495 ± 37 48 ± 6.9 32 ± 4.8

As shown in the table 2, it is obvious that the yield of monacolin K inred mold dioscorea is 5.27 times more than that in red mold rice, 4.95times more than that in cassava, 13.18 times more than that in sweetpotato, and 5.22 times more than that in potato. Thus, dioscorea is thebest substrate for producing monacolin K in these substrates.Furthermore, the table 2 also indicates that the yield of yellow pigmentin red mold dioscorea is higher than that in other substrates, hence redmold dioscorea indeed can promote the yield of yellow pigment, such asmonascin and ankaflavin.

From the above researches, the results demonstrate that red molddioscorea possesses higher yields of monascin, ankaflavin, GABA, andmonacolin K, which are active ingredients with the effects ofanti-inflammation, anti-cancer, blood pressure lowering, nerveconduction promoting, and cholesterol lowering, and can achieve welleffects of blood lipid lowering, blood pressure lowering,atherosclerosis preventing, and Alzheimer's disease improving, thus redmold dioscorea is highly important in the researches and developments ofhealth foods in future. The present invention opens up and develops redmold dioscorea possessing higher yields of functional metabolitescomparing with that in red mold rice, and it is expected to becontributive to the developments of Monascus related products.

The foregoing embodiments are provided to illustrate and disclose thetechnical principles and features of the present invention so as toenable persons skilled in the art to understand the disclosure of thepresent invention and implement the present invention accordingly, andare not intended to be restrictive of the scope of the presentinvention. Hence, all equivalent modifications and variations made tothe foregoing embodiments without departing from the spirit andprinciples in the disclosure of the present invention should fall withinthe scope of the invention as set forth in the appended claims.

What is claimed is:
 1. A method for manufacturing red mold dioscoreacomprising the steps of: (1a) adding a specific amount of water to a rawmaterial of a dioscorea to make the raw material of the dioscorea andthe water be a specific ratio, wherein the raw material of the dioscoreais a dried dioscorea containing sulfur, and the sulfur contained by thedried dioscorea may contribute an acidic condition with pH 3-4, and theacidic condition being able to simulate the production of secondarymetabolites of the red mold dioscorea when the red mold dioscorea iscultured; (2a) proceeding a sterilization process to the raw material ofthe dioscorea and then cooling the product down to a specifictemperature; (3a) inoculating a Monascus spp. to the sterilizeddioscorea; (4a) culturing the inoculated dioscorea under a specificcultural temperature and a specific shaking frequency for a specificcultural time period; (5a) proceeding an anaerobic treatment to theproduct of step (4a) for a specific treating time period, wherein theanaerobic treatment is carried out by culturing the inoculated dioscoreaunder 100% CO₂ condition; and (6a) drying the product of step (5a) to aspecific water content, wherein the specific water content is below 15%.2. The method for manufacturing red mold dioscorea according to claim 1,wherein a water content of the dried dioscorea containing sulfur isbelow 15%, and a sulfur content is below 160 ppm.
 3. The method formanufacturing red mold dioscorea according to claim 1, wherein thespecific ratio in the step (1a) is ranged from 1:10 to 1:200.
 4. Themethod for manufacturing red mold dioscorea according to claim 1,wherein the sterilization process in the step (2a) is carried out at121° C. for 30 minutes.
 5. The method for manufacturing red molddioscorea according to claim 1, wherein the specific culturaltemperature in the step (4a) is 25˜37° C., the specific shakingfrequency is 50˜300 rpm, and the specific cultural time period is 8˜20days.
 6. The method for manufacturing red mold dioscorea according toclaim 1, wherein the specific treating time period in the step (5a) is48 hr.
 7. The method for manufacturing red mold dioscorea according toclaim 1, wherein the product of the step (5a) can be treated with acentrifugation process before going to step (6a).